Based on recent success in cultivation of human epidermal keratinocytes with lethally irradiated mouse 3T3 feeder layers, this project is designed to develop a model system for the study of primate vaginal and cervical epithelial cells in serial culture: 1. Vaginal and ectocervical epithelial cells from normal biopsy tissue will be examined in culture for growth and biochemical properties. First, culture conditions will be optimized for growth rate and differentiated character by variation of epidermal growth factor, cholera toxin, and hydrocortisone concentrations. The density and cell type (3T3 or stromal) or the feeder layer will also be varied. Second, the biochemical markers of the cells (keratins, cross-linked envelopes, glycogen) will be examined to determine the resemblance of the cells to their counterparts in vivo and to epidermal cells. Third, the response of these cells to sex steroid hormones will be studied. The presence of estrogen and progesterone receptors will be examined. 2. The cells from lesions of cervical dysplasia and carcinoma will be cultivated and compared in properties to normal cells by the criteria of growth and differentiated character. The objective is to isolate abnormal cells in culture that are at intermediate stages of transformation. A possible correlation of deficient cross-linked envelope formation with degree of abnormality will be explored. Epithelial cells from biopsy of endocervix will be examined as a necessary control for identification of abnormal cells and for extension of the culture system to pathological conditions such as diethylstilbestrol-induced vaginal adenosis. A long range goal of this research is to find a source of partly transformed human epithelial cells for use as sensitive targets in testing of chemical carcinogens.